Anti-TRAF6 Antibody Picoband™

Product Name

Anti-TRAF6 Antibody Picoband™

View all TRAF-6 Antibodies

SKU/Catalog Number

A00185

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-TRAF6 Antibody Picoband™ catalog # A00185. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-TRAF6 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00185)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived mouse TRAF6 recombinant protein (Position: Q14-D512).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A00185 is reactive to TRAF6 in Mouse, Rat

Applications

A00185 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

60-65 kDa

Calculated molecular weight

59.573kDa

Background of TRAF-6

TNF receptor-associated factor 6, also called E3 ubiquitin-protein ligase TRAF6 or RNF85, is a TRAF human protein. The protein encoded by this gene is a member of the TNF receptor associated factor (TRAF) protein family. TRAF proteins are associated with, and mediate signal transduction from, members of the TNF receptor superfamily. This gene is mapped to 11p12. This protein mediates signaling from members of the TNF receptor superfamily as well as the Toll/IL-1 family. Signals from receptors such as CD40, TNFSF11/RANCE and IL-1 have been shown to be mediated by this protein. This protein also interacts with various protein kinases including IRAK1/IRAK, SRC and PKCzeta, which provides a link between distinct signaling pathways. This protein functions as a signal transducer in the NF-kappaB pathway that activates IkappaB kinase (IKK) in response to proinflammatory cytokines. The interaction of this protein with UBE2N/UBC13, and UBE2V1/UEV1A, which are ubiquitin conjugating enzymes catalyzing the formation of polyubiquitin chains, has been found to be required for IKK activation by this protein.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

"Western blot, 0.25-0.5μg/ml, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat
Flow Cytometry, 1-3μg/1x10⁶ cells, Mouse
Direct ELISA, 0.1-0.5μg/ml, Mouse
"

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of TRAF6 using anti-TRAF6 antibody (A00185).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat RH35 whole cell lysates,
Lane 2: mouse thymus tissue lysates,
Lane 3: mouse spleen tissue lysates,
Lane 4: mouse RAW246.7 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-TRAF6 antigen affinity purified polyclonal antibody (Catalog # A00185) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for TRAF6 at approximately 60-65KD. The expected band size for TRAF6 is at 60KD.

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Figure 2. IHC analysis of TRAF6 using anti-TRAF6 antibody (A00185).
TRAF6 was detected in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-TRAF6 Antibody (A00185) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of TRAF6 using anti-TRAF6 antibody (A00185).
TRAF6 was detected in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-TRAF6 Antibody (A00185) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of HEPA 1-6 cells using anti-TRAF6 antibody (A00185).
Overlay histogram showing HEPA 1-6 cells stained with A00185 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TRAF6 Antibody (A00185, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For TRAF6 (Source: Uniprot.org, NCBI)

Alternative Names

E3 ubiquitin-protein ligase TRAF6; EC 6.3.2.-; Interleukin-1 signal transducer; RING finger protein 85; RNF85MGC:3310; TNF receptor-associated factor 6; TRAF6; TRAF-6 TRAF6 MGC:3310, RNF85 TNF receptor associated factor 6 TNF receptor-associated factor 6|E3 ubiquitin-protein ligase TRAF6|RING finger protein 85|RING-type E3 ubiquitin transferase TRAF6|TNF receptor-associated factor 6, E3 ubiquitin protein ligase|interleukin-1 signal transducer

For more info on TRAF6, check out the TRAF6 Infographic

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In this infographic, you will see the following information for TRAF6: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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